• Journal of Korean Society of Environmental Engineers
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• v.33 no.9
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• pp.662-669
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• 2011

This study was conducted to biologically convert methane into methanol. Methane contained in biogas was bio-catalytically oxidized by methane monooxygenase (MMO) of methanotrophs, while methanol conversion was observed by inhibiting methanol dehydrogenase (MDH) using MDH activity inhibitors such as phosphate, NaCl, $NH_4Cl$, and EDTA. The degree of methane oxidation by methanotrophs was the most highly accomplished as 0.56 mmol for the condition at $35^{\circ}C$ and pH 7 under 0.4 (v/v%) of biogas ($CH_4$ 50%, $CO_2$ 50%) / Air ratio. By the inhibition of 40 mM of phosphate, 50 mM of NaCl, 40 mM of $NH_4Cl$ and $150{\mu}m$ of EDTA, methane oxidation rate could achieve more than 80% regardless of type of inhibitors. In the meantime, addition of 40 mM of phosphate, 100 mM of NaCl, 40 mM of $NH_4Cl$ and $50{\mu}m$ of EDTA each led to generating the highest amount of methanol, i.e, 0.71, 0.60, 0.66, and 0.66 mmol when 1.3, 0.67, 0.74, and 1.3 mmol of methane was each concurrently consumed. At that time, methanol conversion rate was 54.7, 89.9, 89.6, and 47.8% respectively, and maximum methanol production rate was $7.4{\mu}mol/mg{\cdot}h$. From this, it was decided that the methanol production could be maximized as 89.9% when MDH activity was specifically inhibited into the typical level of 35% for the inhibitor of concern.

• KSBB Journal
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• v.8 no.4
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• pp.341-350
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• 1993

As an effort to develop an alternative transportation fuel, the production of methanol from methane gas was studied using the resting cells of an obligatory methanotroph, Methylosinus trichosporium OB3b. The reaction was carried out in high concentration phosphate buffer solutions with the flask-grown cells containing the exclusively cytoplasmic methane monooxygenase (sMMO) activity. The methanol accumulation rate was observed to be 79nmo1/mg·min during the initial 4.5hr. Phosphate-dependent inhibition was found for both sMMO and methanol dehydrogenase (MDH) activities, and the inhibition constants were 185mM and 42mM, respectively. The inhibition mode was noncompetitive. Methanol was found to be very inhibitory to the sMMO activity and the inhibition constant (noncompetitive) was 21mM when propylene was used as substrate. The sMO activity in the resting cells was declined very fast and the rate became very high during the methanol production. These results indicate that the use of M. trichosporium OB3b as a biocatalyst for the methanol production is heavily dependent on the stable maintenance of the whole-cell SMO activity as well as the effective alleviation of product inhibition.

• YAKHAK HOEJI
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• v.34 no.6
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• pp.447-456
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• 1990

Various methods are available for the production of L-threonine. The microbial production of L-threonine has been achieved by breeding L-threonine analog-resistant auxotrophic mutants of various bacteria. The enzymatic production of L-threonine has been demonstrated by use of threonine metabolic enzymes such as threonine deaminase, threonine aldolase, or threonine dehydrogenase complex. Threonine synthesis from glycine and ethanol seems to be catalyzed by the enzymes Methanol dehydrogenase(MDH) and Serine hydroxymethyltransferase(SHMT), which was also found to catalyze the aldol condensation of glycine with acetaldehyde. The improved production of L-threonine has been achieved by amplifying the genes for the L-threonine biosynthetic enzymes using recombinant DNA techniques.

• KSBB Journal
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• v.10 no.2
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• pp.212-220
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• 1995

An obligatory type II methanotroph Methylosinus trichosporium OB3b was cultivated on methanol as a sole carbon and energy source. The effects of methanol concentration, pH, temperature, nitrogen source and phosphate concentration on cell growth were investigated and the results were compared with the growth on methane, which had been studied previously. When $(NH_4)_2SO_4$ was used as a nitrogen source, the maximal specific growth rate (${\mu}max$) on methanol was $0.20hr^{-1}$ and the carbon conversion efficiency(CCE) was 43%. In comparison, on methane, ${\mu}max$ and CCE were $0.08hr^{-1}$ and 32%, respectively. Ammonia was found to be a better nitrogen source for methanol-growing cells. Cell yield on nitrogen (YX/N) was the same regardless of nitrogen source as 7.14g dry cells/g N, but the yield on methanol(YX/N) was higher with ammonia(0.8g dry cells/g MeOH) than with nitrate(0.64g dry cells/g MeOH). Optimal pH and temperature were 7.0 and $30^{\circ}C$, respectively. Methanol inhibition on cell growth was observed at above 0.5%(v/v). Inhibition by phosphate was observed at above 60mM, although the inhibition on methanol dehydrogenase activity started at a much lower level of 20mM. Based on the experimental findings, the cellular physiology of M. trichosporium OB3b growing on the two closely-related carbon sources were discussed extensively.

• Proceedings of the Zoological Society Korea Conference
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• 2001.10a
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• pp.244.3-245
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• 2001

No Abstract, See Full Text

• KSBB Journal
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• v.32 no.2
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• pp.124-132
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• 2017

A methane-oxidizing bacterium was isolated from rice paddy field soil around Jeollanam-do province, Korea, and characterized. The isolate was gram-negative, orange pigmented and short rod ($1.1-1.2{\times}1.6-1.9{\mu}m$). It was catalase and urease-negative but oxidase-positive. The strain utilized methane and methanol as sole carbon and energy sources. It had an ability to grow with an optimum pH 7.0 and an optimum growth temperature $30^{\circ}C$. The strain was resistant to antibiotic polymyxin B but sensitive to streptomycin, kanamycin, ampicillin, chloramphenicol and rifampicin. The isolate required copper for their growth with concentration range of $2-25{\mu}M$, with an optimum of $10{\mu}M$. Under optimal culture condition, specific cell growth rate and generation time were found to be $0.046hr^{-1}$ and 15.13 hr, respectively. Phylogenetic analysis based on 16S rDNA sequences indicated that the strain formed a tight phylogenetic lineage with Methylomonas koyamae with a value of 99.4% gene sequence homology. So, we named the isolate as Methylomonas sp. SM4. 8.6 mM methanol was accumulated in the reaction mixture containing 70 mM sodium formate and 40 mM $MgCl_2$ (MDH inhibitor) under atmosphere of methane:air (40:60) mixture for 24 hr at $30^{\circ}C$.